The acclimation of plants to moderately high temperature plays an important role in inducing plant tolerance to subsequent lethal high temperatures. This study was performed to investigate the effects of heat acclimation and sudden heat stress on protein synthesis and degradation in creeping bentgrass (Agrostis palustris Huds.). Plants of the cultivar Penncross were subjected to two temperature regimes in growth chambers: 1) heat acclimation—plants were exposed to a gradual increase in temperatures from 20 to 25, 30, and 35 °C for 7 days at each temperature level before being exposed to 40 °C for 28 days; and 2) sudden heat stress (nonacclimation)—plants were directly exposed to 40 °C for 28 days from 20 °C without acclimation through the gradual increase in temperatures. Heat acclimation increased plant tolerance to subsequent heat stress, as demonstrated by lower electrolyte leakage (relative EL) in leaves of heat-acclimated plants compared to nonacclimated plants at 40 °C. Heat acclimation induced expression of some heat shock proteins (HSPs), 57 and 54 kDa, detected in a salt-soluble form (cystoplasmic proteins), which were not present in unacclimated plants under heat stress. However, HSPs of 23, 36, and 66 kDa were induced by both sudden and gradual exposure to heat stress. In general, total protein content decreased under both heat acclimation and sudden heat stress. Cystoplasmic proteins was more sensitive to increasing temperatures, with a significant decline initiated at 25 °C, while sodium dodecyl sulphate (SDS)-soluble (membrane) protein content did not decrease significantly until temperature was elevated to 30 °C. The results demonstrated that both a gradual increase in temperature and sudden heat stress caused protein degradation and also induced expression of newly synthesized HSPs. Our results suggested that the induction of new HSPs during heat acclimation might be associated with the enhanced thermotolerance of creeping bentgrass, although direct correlation of these two factors is yet to be determined. This study also indicated that protein degradation could be associated with heat injury during either gradual increases in temperature or sudden heat stress.
Yali He, Xiaozhong Liu and Bingru Huang
Chenping Xu, Zhongchun Jiang and Bingru Huang
Nitrogen (N) deficiency inhibits plant growth and induces leaf senescence through regulating various metabolic processes. The objectives of this study were to examine protein changes in response to N deficiency in immature and mature leaves of a perennial grass species and determine major metabolic processes affected by N deficiency through proteomic profiling. Creeping bentgrass (Agrostis stolonifera cv. Penncross) plants were originally fertilized with a diluted 36N–2.6P–5K fertilizer. After 14 days acclimation in a growth chamber, plants were grown in a nutrient solution containing 6 mm nitrate (control) or without N (N deficiency). Immature leaves (upper first and second not yet fully expanded leaves) and mature leaves (lower fully expanded leaves) were separated at 28 days of treatment for protein analysis. Two-dimensional electrophoresis and mass spectrometry analysis were used to identify protein changes in immature and mature leaves in response to N deficiency. The abundance of many proteins in both immature and mature leaves decreased with N deficiency, including those involved in photosynthesis, photorespiration, and amino acid metabolism (hydroxypyruvate reductase, serine hydroxymethyltransferase, alanine aminotransferase, glycine decarboxylase complex, glycolate oxidase), protein protection [heat shock protein (HSP)/HSP 70, chaperonin 60 and FtsH-like protein], and RNA stability (RNA binding protein). The reduction in protein abundance under N deficiency was greater in mature leaves than in immature leaves. The abundance of small HSP and metalloendopeptidase increased under N deficiency only in immature leaves. These results suggest that N deficiency accelerated protein degradation in immature and mature leaves of creeping bentgrass, particularly those proteins associated with energy and metabolism, but to a lesser extent in immature leaves. Immature leaves were also able to accumulate proteins with chaperone functions and for N reutilization, which could protect leaves from senescence under N deficiency.
Eric Watkins, Bingru Huang and William A. Meyer
Tufted hairgrass [Deschampsia cespitosa (L.) Beauv.] is receiving increasing attention as a low-maintenance turfgrass for use in areas with reduced fertility or reduced sunlight. The objectives of this study were to examine physiological responses of tufted hairgrass to heat and drought stress and to distinguish whether better summer performance was related to better heat or drought tolerance. Four germplasm lines were chosen based on summer performance in field plots (two lines resistant to summer stress and two lines susceptible to summer stress) and were grown in growth chambers [14-hour photoperiod, 20/15 °C (day/night)]. Plants were exposed to either drought stress or heat stress (35/30 °C, day/night) for up to 49 days. Control plants maintained under normal conditions (20/15 °C, day/night, well watered) were included for both treatments. During the course of the study, single-leaf photosynthetic rate, photochemical efficiency, and relative water content were measured, and turf quality was visually rated. All parameters for all tufted hairgrass lines decreased under drought stress and heat stress, and the decline was more severe for summer stress-susceptible lines than for resistant lines. Lines that were previously considered resistant to summer stress exhibited superior photochemical efficiency under heat stress compared with the susceptible lines. When subjected to drought stress, the lines exhibited little or no differences in the measured parameters. These results suggest that observed variation in field summer performance among various tufted hairgrass germplasm lines may be mainly the result of their differences in heat tolerance. These results suggest that selecting for heat-tolerant germplasm could be important for further improvement in turf performance of tufted hairgrass during the summer.
Mahalaxmi Veerasamy, Yali He and Bingru Huang
Heat stress induces leaf senescence and causes changes in protein metabolism. The objective of this study was to investigate effects of exogenous application of a synthetic form of cytokinin, zeatin riboside (ZR), on protein metabolism associated with leaf senescence under heat stress for a cool-season grass species. Creeping bentgrass (Agrostis stolonifera L.) (cv. Penncross) plants were exposed to optimum temperature control (20/15 °C, day/night) and heat stress (35/30 °C) in growth chambers. Before heat stress treatments, foliage was sprayed with 10 μmol ZR or water (untreated) for 3 days and then once per week during 35 days of heat stress. Leaf chlorophyll content, photochemical efficiency (Fv/Fm), and soluble protein content declined, whereas protease activity increased during heat stress. Treatments with ZR helped maintain higher leaf chlorophyll content, Fv/Fm, and soluble protein content under heat stress. Protease activity in ZR-treated plants was lower than that of untreated plants. Zeatin riboside-treated plants had less severe degradation of ribulose-1,5-bisphosphate carboxylase proteins than untreated plants exposed to heat stress. In addition, ZR treatment upregulated the expression of 32- and 57-kDa proteins under heat stress conditions. These results demonstrated that the exogenous application of ZR ameliorated the negative effects of heat stress, as manifested by suppression or delay of leaf senescence. Cytokinins may have helped to alleviate heat stress injury, probably by slowing down the action of protease and by induction or upregulation of heat-shock proteins.
Jinmin Fu, Jack Fry and Bingru Huang
Deficit irrigation is increasingly used to conserve water, but its impact on turfgrass rooting has not been well documented. The objective of this study was to examine the effects of deficit irrigation on ‘Falcon II’ tall fescue (Festuca arundinacea Schreb.) root characteristics in the field using a minirhizotron imaging system. The experiment was conducted on a silt loam soil from the first week of June to mid-Sept. 2001 and 2002 using a mobile rainout shelter under which turf received applications of 20%, 60%, or 100% of actual evapotranspiration (ET) twice weekly. Neither soil water content (0 to 25 cm) nor tall fescue rooting between 4.1- and 50.1-cm depths was affected by irrigation at 60% compared with 100% ET. Despite consistently lower soil water content, tall fescue irrigated at 20% ET exhibited an increase in root parameters beginning in July or August. Tall fescue subjected to 20% ET irrigation had greater total root length and surface area on two of five monitoring dates in 2002 compared with that receiving 100% ET. Evaluation of tall fescue rooting by depth indicated that root proliferation at 20% ET was occurring between 8.7- and 36.3-cm depths. As evaluated under the conditions of this experiment, turfgrass managers using deficit irrigation as a water conservation strategy on tall fescue should not be concerned about a reduction in rooting deep in the soil profile, and irrigation at 20% ET may result in root growth enhancement.
Jinmin Fu, Jack Fry and Bingru Huang
Water requirements for `Meyer' zoysiagrass (Zoysia japonica Steud., hereafter referred to as zoysia), `Midlawn' bermudagrass [Cynodon dactylon (L.) Pers. × C. transvaalensis Burtt-Davy, hereafter referred to as bermuda], `Falcon II' tall fescue (Festuca arundinacea Schreb.) and `Brilliant' kentucky bluegrass (Poa pratensis L., hereafter referred to as bluegrass) were evaluated under a mobile rainout shelter at deficit irrigation levels of 20% to 100% of actual evapotranspiration (ETa), applied twice weekly, between June and September 2001 and 2002. Soil was a river-deposited silt loam (fine, montmorillonitic, mesic Aquic Arquidolls). Minimum annual irrigation amounts required to maintain quality ranged from 244 mm for bermuda to 552 mm for bluegrass. Turfgrass species and respective irrigation levels (% of ETa) at which season-long acceptable turf quality was maintained in each year were bluegrass, 100% (evaluated 2001 only); tall fescue, 60% in 2001 and 80% in 2002; bermuda, 60% in both years; and zoysia, 80% in both years. A landscape manager who could tolerate one week of less-than-acceptable quality could have irrigated tall fescue at 40% ETa (224 mm) in 2001 and 60% ETa (359 mm) in 2002. Likewise, bermuda exhibited unacceptable quality on only one September rating date when irrigated at 40% ETa (163 mm) in 2001. Bermuda was able to tolerate a lower leaf relative water content (LRWC) and higher level of leaf electrolyte leakage (EL) compared to other grasses before quality declined to an unacceptable level.
Qingzhang Xu, Bingru Huang and Zhaolong Wang
Turf quality of creeping bentgrass (Agrotis palustris L.) often declines during summer months. Reducing soil temperature alleviates bentgrass quality decline at supraoptimal air temperatures. The objective of this study was to investigate whether reducing soil temperature during the night is more effective than during the day in improving shoot and root growth when air temperature was supraoptimal for creeping bentgrass. The experiment was conducted in growth chambers using water baths to manipulate soil temperatures. Plants were exposed to the following temperature treatments: 1) optimal air and soil temperature during the day and night (20/20 °C, day/night, control); 2) high air and soil temperature during the day and night (35/35 °C, day/night); 3) lower soil temperatures during the day (20/35, 25/35, and 30/35 °C, day/night); and 4) lower soil temperature during the night (35/20, 35/25, and 35/30 °C) while air temperature was maintained at 35 °C during the day and night. Turf quality (on 1-9 scale) increased to the level of 6.5, 3.0, and 2.5 by reducing day soil temperature to 20, 25, or 30 °C, respectively, at 28 days of treatment, compared to the quality of 2.0 at 35/35 °C. Turf quality increased from 2.0 at 35/35 °C to 7.0, 6.0, and 4.5, respectively, by 28 days of exposure to night temperatures of 20, 25, and 30 °C. Chlorophyll content, root number, and root weight also were increased by reducing day or night soil temperature, and the increases were more pronounced for reduced night temperatures than day temperatures. These results demonstrated that reduced night soil temperature was more effective than reduced day soil temperature in improving shoot and root growth in creeping bentgrass under high air temperature conditions.
Bingru Huang, Jack Fry and Bin Wang
Understanding factors associated with drought resistance and recovery from drought stress in tall fescue (Festuca arundinaces Schreb.) is important for developing resistant cultivars and effective management strategies. Our objective was to investigate water relations, photosynthetic efficiency, and canopy characteristics of tall fescue cultivars (forage-type `Kentucky-31', turf-type `Mustang', and dwarf-type `MIC18') in responses to drought stress and subsequent recovery in the field and greenhouse. During drought stress under field conditions, `MIC18' had lower turf quality, more severe leaf wilting, and higher canopy temperature than `Mustang' and `Kentucky-31', indicating that `MIC18' was more drought-sensitive. The greenhouse study comparing `K-31' and `MIC18' showed that leaf water status, chlorophyll fluorescence, canopy green leaf biomass, and lead area index of both cultivars declined as soil dried. Reductions in relative water content, leaf water potential, chlorophyll fluorescence, canopy green leaf biomass, and leaf area index were more severe and occurred sooner during dry down for `MIC18' than for `Kentucky-31'. After rewatering following 14 days of stress, leaf water deficit and turf growth recovered, to a greater degree for `Kentucky-31' than for `MIC18'. However, soil drying for 21 days caused long-term negative effects on leaf photosynthetic efficiency and canopy characteristics for both cultivars.
Zhaolong Wang, John Pote and Bingru Huang
This study was designed to examine whether shoot injury induced by high root-zone temperature is associated with changes in shoot detoxifying metabolism and to determine the level and duration of high root-zone temperatures that would induce physiological changes in two cultivars of creeping bentgrass (Agrostis stolonifera var. palustris Huds) differing in heat tolerance. Plants of `Penn A-4' (heat tolerant) and `Putter' (heat susceptible) were grown in sand and exposed to root-zone temperatures of 20 (control), 21, 22, 23, 25, 27, 31, and 35 °C in water baths while air temperature was maintained at 20 °C in a growth chamber. Turf quality, leaf cytokinin content, and antioxidant enzyme activities declined at increased soil temperatures and the duration of treatment for both cultivars. A decline in turf quality occurred following 40 days of exposure to 35 °C for `Penn A-4' and 26 days of exposure to 31 °C for `Putter'. The root-zone temperature causing the decline of isopentenyl adenosine and zeatin cytokinins was 25 °C at 37 d for `Putter' and 27 °C at 47 days for `Penn A-4'. The temperature causing the decline of superoxide dismutase and catalase activities was 25 °C and 27 °C at 33 days for `Putter' and 27 °C and 31 °C at 43 days for Penn A-4, respectively. Malondialdehyde content increased at 27 °C for `Putter' and 31 °C for `Penn A-4' at 43 days of treatment. The decline in cytokinin content and antioxidant enzyme activity occurred at a lower soil temperature and earlier during the treatment than the decline in turf quality, possibly contributing to turf quality decline. The root-zone temperatures causing the decline in turf quality, cytokinin content, and oxidative damage were higher in the heat-tolerant cultivar than heat-susceptible cultivar.
Qingzhang Xu, Bingru Huang and Zhaolong Wang
Heat injury in creeping bentgrass (Agrostis stolonifera var. palustris Huds) has been associated with decreases in carbohydrate availability. Extending light duration may increase carbohydrate availability and thus improve growth of creeping bentgrass under heat stress. The objective of this study was to investigate whether turf performance and carbohydrate status could be improved by extending daily light duration for creeping bentgrass exposed to supraoptimal temperature conditions. `Penncross' plants were initially grown in growth chambers set at a day/night temperature of 20/15 °C and 14-hour photoperiod and then exposed to a day/night temperature of 33/28 °C (heat stress) and three different light durations: 14 (control), 18, and 22 hours (extended light duration) for 30 days. Turf quality and tiller density decreased with the duration of heat stress, as compared to the initial level at 20 °C, regardless of the light duration. However, both parameters increased with extended light duration from 14 to 18 or 22 hours. Extended light duration, particularly to 22 hours, also improved canopy net photosynthetic rate from -1.26 to 0.39 μmol·m-2·s-1 and daily total amount of carbon assimilation from -6.4 to 31.0 mmol·m-2·d-1, but reduced daily total amount of carbon loss or consumption to 50% through dark respiration compared to 14 hours treatment by the end of experiment. In addition, extending light duration from 14 to 22 hours increased water-soluble carbohydrate content in leaves both at the end of light duration and the dark period. These results demonstrated that extending light duration improved turf performance of creeping bentgrass under heat stress, as manifested by the increased tiller density and turf quality. This could be related to the increased carbohydrate production and accumulation. Supplemental lighting could be used to improve performance if creeping bentgrass is suffering from heat stress.